Specific hrp dab detection ihc kit

The suspensions of CTC, SOX9-TC, IGF-I-TC, and NTC
were adjusted to densities of 2×10⁴ cells/mL in complete medium, and 500
µL aliquots were seeded into each compartment of a four-well microchamber slide
(NuncTM; Thermo Fisher Scientific). One microchamber slide was prepared for each
transfection condition, and the microchamber slides were immediately incubated at
37°C in a 5% CO₂ atmosphere for 3, 6, or 9 days. Next, the culture medium
was removed and the cells were fixed with methanol-acetone (1:1 v/v) for 20 min at
-20°C. The NTC, IGF-I-TC, SOX9-TC, and CTC on the
3rd, 6th, and 9th PT days were incubated with monoclonal antibodies against both
Col-I (ab23446; Abcam, Inc., USA) and Col-II (ab34712; Abcam, Inc.), and positive
staining was detected using mouse- and rabbit-specific HRP/DAB detection IHC Kit
(Abcam, Inc.), according to the manufacturer's instructions.

Streptozotocin (STZ) and curcumin were purchased from Sigma-Aldrich Inc., St. Louis, MO, USA. The antioxidant enzyme kits including catalase, glutathione, and superoxide dismutase were acquired from Abcam, Cambridge, UK. The inflammatory markers kits for TNF-α, IL-6 and IL-1β were also obtained from Abcam, UK. Antioxidant enzyme and myeloperoxidase kits were purchased from Abcam, UK. For fibrosis evaluation, trichrome stain and a Sirius red kit were purchased from the same company. IL-6 and TNF-α monoclonal antibodies and the Specific HRP/DAB Detection IHC kit were acquired from Abcam, United Kingdom. All supportive chemicals used in this study were of analytical grade and purchased from a local vendor.